SUBSTITUTED BENZO[5,6][1,4]DIAZEPINO[1,2-a]INDOLES FOR THE TREATMENT OF PROLIFERATIVE DISORDERS

Abstract

The invention relates to novel benzodiazepine derivatives with antiproliferative activity and more specifically to novel benzodiazepine compounds of formulae (I), (II) and (III). The invention also provides conjugates of the benzodiazepine compounds linked to a cell-binding agent. The invention further provides compositions and methods useful for inhibiting abnormal cell growth or treating a proliferative disorder in a mammal using the compounds or conjugates of the invention.

1 - 81 . (canceled) 
     
     
         82 . A cell-binding agent-cytotoxic agent conjugate represented by the following formula: 
       
         
           
           
               
               
           
         
       
       or a pharmaceutically acceptable salt thereof, wherein:
 W 1  is —(CH 2 ) n1 —; 
 W 2  is —(CH 2 ) n2 — 
 n 1  is 1, 2 or 3; 
 n 2  is 0, or 2; 
 n 3  is 0 or 1; 
 R 1a  and R 1a′  are each independently H, halide, —OH, or (C 1 -C 6 )alkyl; or R 1a  and R 1a′  together form a double bond containing group ═B; 
 ═B is selected from a (C 2 -C 6 )alkenyl or a carbonyl group, wherein the (C 2 -C 6 )alkenyl is optionally substituted with a halogen, —OH, (C 1 -C 3 )alkoxy or phenyl; 
 Q is Q 1 -Ar-Q 2 ; 
 Q 1  is absent, (C 1 -C 6 )alkyl, or —CH═CH—; 
 Ar is absent or an aryl group; 
 Q 2  is —H, a (C 1 -C 6 )alkyl, a (C 1 -C 6 )alkenyl, a polyethylene glycol unit —R c′ —(OCH 2 CH 2 ) n —R c , or a substituent selected from a halogen, guanidinium [—NH(C═NH)NH 2 ], —R, —OR, —NR′R″, —NO 2 , —NCO, —NR′COR″, NR′(C═O)OR″—SR, —SOR′, —SO 2 R′, —SO 3 H, —OSO 3 H, —SO 2 NR′R″, cyano, an azido, —COR′, —OCOR′ and —OCONR′R″; 
 n is an integer from 1 to 10; 
 R c′  is a (C 1 -C 4 )alkyl, 
 R c  is H, or a (C 1 -C 4 )alkyl, 
 the double line  between N and C represents a single bond or a double bond, provided that when it is a double bond X is absent and Y is —H, or a (C 1 -C 4 )alkyl, and when it is a single bond, X is —H, an amine protecting moiety or R L1 ; and 
 Y is H of a leaving group selected from —OR Y , —OCOR Y1 , —OCOOR Y1 , —OCONR Y1 R Y2 , —NR Y1 R Y2 , —NR Y1 COR Y2 , —NR Y1 NR Y1 R Y2 , an optionally substituted 5- or 6-membered nitrogen-containing heterocycle (e.g., piperidine, tetrahydropyrrole, pyrazole, morpholine, etc. attached through the nitrogen atom), a guanidinium represented by —NR Y1 (C═NH)NR Y1 R Y2 , an amino acid residue, or a peptide represented by —NRCOP′, —SR Y , —SOR Y1 , halogen, cyano, azido, —OSO 3 H, sulfite (—SO 3 H or —SO 2 H), metabisulfite (H 2 S 2 O 5 ), mono-, di-, tri-, and tetra-thiophosphate (PO 3 SH 3 , PO 2 S 2 H 2 , POS 3 H 2 , PS 2 H 2 ), thiophosphate ester (R i O) 2 PS(OR i ), R i S—, R i SO, R i SO 2 , R i SO 3 , thiosulfate (HS 2 O 3 ), dithionite (HS 2 O 4 ) phosphorodithioate (P(═S)(OR k′ )(S)(OH)), hydroxamic acid (R k′ C(═O)NOH), and formaldehyde sulfoxylate (HOCH 2 SO 2 —) or a mixture thereof, wherein R i  is a linear or branched alkyl having 1 to 10 carbon atoms and is substituted with at least one substituent selected from —N(R j ) 2 , —CO 2 H, —SO 3 H, and —PO 3 H; R i  can be further optionally substituted with a substituent for an alkyl described herein; R j  is a linear or branched alkyl having 1 to 6 carbon atoms; R k′  is a linear, branched or cyclic alkyl, alkenyl or alkynyl having 1 to 10 carbon atoms, aryl, heterocyclyl or heteroaryl; 
 P′ is an amino acid residue or a polypeptide containing between 2 to 20 amino acid residues, 
 R Y , for each occurrence, is independently selected from the group consisting of —H, an optionally substituted linear, branched or cyclic alkyl, alkenyl or alkynyl having from 1 to 10 carbon atoms, a polyethylene glycol unit —(CH 2 CH 2 O) n —R c , an optionally substituted aryl having 6 to 18 carbon atoms, an optionally substituted 5- to 18-membered heteroaryl ring containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur, or an optionally substituted 3- to 18-membered heterocyclic ring containing 1 to 6 heteroatoms independently selected from O, S, N and P; 
 R Y1  and R Y2  are each independently selected from —H, —OH, —OR Y , —NHR Y , —NR Y   2 , —COR Y , an optionally substituted linear, branched or cyclic alkyl, alkenyl or alkynyl having from 1 to 10 carbon atoms, a polyethylene glycol unit; 
 R L1  is self-immolative linker bearing a linking moiety that is covalently bonded the cell-binding agent (CBA); 
 R 1 , R 2 , R 3  and R 4  are each independently selected from —H, a (C 1 -C 6 )alkyl, halogen, —OR, —NR′R″, —NO 2 , —NR′COR″, —SR, —SOR′, —SO 2 R′, —SO 3 H, —OSO 3 H, —SO 2 NR′R″, cyano, —COR′, —OCOR′, and —OCONR′R″; 
 R, for each occurrence, is —H or a (C 1 -C 6 )alkyl; 
 R′ and R″ are each independently selected from —H, —OH, —OR, —NHR, —NR 2 , —COR, or a (C 1 -C 6 )alkyl; 
 R 6  is —H, —R, —OR, —SR, —NR′R″, —NO 2 , or halogen; 
 A is absent or is selected from —O—, —C(═O)—, —CRR′O—, —CRR′—, —S—, —CRR′S—, —NR 5  and —CRR′N(R 5 )—; 
 L 1  is a spacer, 
 Z 1  is a bond, 
 
       
         
           
           
               
               
           
         
         s3 is the site connected L 1  and s4 is the site connected to Ar 1 ; 
         R 5  is —H or a (C 1 -C 4 )alkyl; 
         R 100  and R 101 , for each occurrence, are each independently —H, halogen, (C 1 -C 4 )alkyl, or (C 1 -C 4 )haloalkyl; 
         Ar 1  is a 6- to 18-membered aromatic ring, a 5- to 18-membered heteroaromatic ring, or —Ar 1 ′—Ar 1 ″—, wherein Ar 1 ′ and Ar 1 ″ are each independently a 6- to 18-membered aromatic ring or a 5- to 18-membered heteroaromatic ring; 
         Z 2  is 
       
       
         
           
           
               
               
           
         
         s5 is the site connected Ar 1  and s6 is the site connected to Ar 2 ; 
         Ar 2  is a 6- to 18-membered aromatic ring, a 5- to 18-membered heteroaromatic ring or —Ar 2 ′—Ar 2 ″—, wherein Ar 2 ′ and Ar 2 ″ are each independently a 6- to 18-membered aromatic ring or a 5 5- to 18-membered heteroaromatic ring; 
       
       
         
           
           
               
               
           
         
         s7 is the site connected to Ar 2  and s8 is the site connected to Ar 3 ; 
         Ar 3  is a 6- to 18-membered aromatic ring, a 5- to 18-membered heteroaromatic ring or —Ar 3 ′—Ar 3 ′—, wherein Ar 3 ′ and Ar 3 ″ are each independently a 6- to 18-membered aromatic ring or a 5-to 18-membered heteroaromatic ring; 
         L CB  is a linker bearing a linking moiety that is covalently linked to the cell-binding agent (CBA); 
         w is an integer from 1 to 20; 
         R a  is —OH, —Cl, —O(C 1 -C 6 )alkyl or —C(═O)OR a  is a reactive ester group; 
         R b  and R c  are each independently —H, (C 1 -C 4 )alkyl or an amine protecting group; and provided (i) when the conjugate is represented by formula (V) or (VI), Z 1  is not 
       
       
         
           
           
               
               
           
         
         (ii) when the conjugate is represented by formula (IV), the conjugate does not comprise a compound represented by 
       
       
         
           
           
               
               
           
         
       
       or a pharmaceutically acceptable salt thereof; and
 (iii) when X is R L1 , L is H, —C(═O)R a  or —NR b R c . 
 
     
     
         83 - 86 . (canceled) 
     
     
         87 . The conjugate of  claim 82 , wherein the conjugate is represented by the following formula: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       or pharmaceutically acceptable salt thereof, wherein
 R 6  is —OR; 
 R is —H or a (C 1 -C 4 )alkyl; 
 A is O; 
 L 1  is —(CH 2 ) m1 —, 
 
       
         
           
           
               
               
           
         
         s1 is the site connected to A; s2 is the site connected to Z 1 ; 
         m1 is 1, 2, 3, 4, 5 or 6; 
         m2 is 1, 2 or 3; 
         m2′ is 0, 1, 2 or 3; and 
         m3 is an integer from 1 to 10. 
       
     
     
         88 - 89 . (canceled) 
     
     
         90 . The conjugate of  claim 87 , wherein R L1  is represented by the following formula: 
       
         
           
           
               
               
           
         
       
       wherein:
 P 1  is a peptide residue having 2 to 5 amino acid residues; and 
 J CB′ -A- is represented by: 
 
       
         
           
           
               
               
           
         
         wherein: s10 is the site connected to the CBA; s9 is the site connected to P 1 ′; m4, m5, m5′, m6, m7 and m8 are each independently an integer from 1 to 6; m5″ and m7′ are each independently 0 or an in from 1 to 10; E is —OH, or —Cl or —C(═O)E is a reactive ester, and J CB   s1  is 
       
       
         
           
           
               
               
           
         
       
       s9 is the site connected to P 1  and s10 is the site connected to CBA through an aldehyde group located on the CBA. 
     
     
         91 - 92 . (canceled) 
     
     
         93 . The conjugate of  claim 82 , wherein L 1  is represented by one of the following formulas: 
       
         
           
           
               
               
           
         
       
       wherein s1 is the site connected to A and s2 is the site connected to Z 1 ; 
       
         
           
           
               
               
           
         
         Z 2  is 
       
       
         
           
           
               
               
           
         
       
       wherein
 R 5  is H or Me. 
 
     
     
         94 . (canceled) 
     
     
         95 . The conjugate of  claim 82 , wherein,
 Ar 1  is benzene, naphthalene, a 5- to 6-membered heteroaromatic ring, a 8 to 10-membered bicyclic heteroaromatic ring or —Ar 1 ′—Ar 1 ″—, wherein Ar 1 ′ and Ar 1 ″ are each independently benzene or a 5- to 6-membered heteroaromatic ring; and   Ar 2  is benzene, naphthalene, naphthalene, a 5- to 6-membered heteroaromatic ring, or a 8- to 10-membered bicyclic heteroaromatic ring, or —Ar 2 ′—Ar 2 ″—, wherein Ar 2 ′ and Ar 2 ″ are each independently benzene or a 5- to 6-membered heteroaromatic ring.   
     
     
         96 - 97 . (canceled) 
     
     
         98 . The conjugate of  claim 95 , wherein Ar 1  is represented by one of the following: 
       
         
           
           
               
               
           
         
       
       wherein R d  is H, a (C 1 -C 6 )alkyl optionally substituted with halogen, —OH, or —NR b R c , wherein R b  and R c  are each independently H, (C 1 -C 4 )alkyl or an amine protecting group, phenyl or heteroaryl; and
 Ar 2  is represented by one of the following: 
 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein R d ′ is H, a (C 1 -C 6 )alkyl optionally substituted with halogen, —OH, or —NR b R c , wherein R b  and R c  are each independently H (C 1 -C 3 )alkyl, an amine protecting group, phenyl or heteroaryl. 
     
     
         99 - 105 . (canceled) 
     
     
         106 . The conjugate of  claim 82 , wherein n 3  is 0. 
     
     
         107 . The conjugate of  claim 82 , wherein n 3  is 1 and Z 3  is 
       
         
           
           
               
               
           
         
       
       wherein R 5  is H or Me. 
     
     
         108 . The conjugate of  claim 107 , wherein Ar 3  is benzene, naphthalene, a 5- to 6-membered heteroaromatic ring, or a 8- to 10-membered bicyclic heteroaromatic ring or —Ar 3 ′—Ar 3 ″—, wherein Ar 3 ′ and Ar 3 ″ are each independently benzene or a 5- to 6-membered heteroaromatic ring. 
     
     
         109 . (canceled) 
     
     
         110 . The conjugate of  claim 108 , wherein Ar 3  is represented by the following formula: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein S8′ is the site connected to Z 3  group; s9′ is the site connected to L CB ; R d ′ is H, a (C 1 -C 6 )alkyl optionally substituted with halogen, —OH, or —NR b R c , wherein R b  and R c  are each independently, (C 1 -C 3 )alkyl, an amine protecting group, phenyl or heteroaryl 
     
     
         111 . The conjugate of  claim 82 , wherein the conjugate is represented by the following formula: 
       
         
           
           
               
               
           
         
       
       or a pharmacuetically acceptable salt thereof, wherein:
 Ar 1  is pyrrole, imidazole, thiazole, pyridine, benzo[b]furan, benzene or —Ar 1 ′—Ar 1 ″—, wherein Ar 1 ′ and Ar 1 ″ are each independently benzene, pyrrole, thiazole, pyrrole; and 
 Ar 2  is pyrrole, imidazole, benzene, benzo[b]thiophene, benzo[b]furan, benzimidazole, indole, quinoline, isoquinoline or —Ar 2 ′—Ar 2 ″—, wherein Ar 2 ′ and Ar 2 ″ are each independently benzene, pyrrole, thiazole, or pyridine. 
 
     
     
         112 . The conjugate of  claim 111 , wherein:
 Ar 1  is   
       
         
           
           
               
               
           
         
       
       wherein R d  is H, a (C 1 -C 6 )alkyl optionally substituted with halogen, —OH, or —NR b R c , wherein R b  and R c  are each independently H, (C 1 -C 4 )alkyl or an amine protecting group, phenyl or heteroaryl; and
 Ar 2  is 
 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein R d ′ is H, a (C 1 -C 6 )alkyl optionally substituted with halogen, —OH, or —NR b R c , wherein R b  and R c  are each independently H, (C 1 -C 3 )alkyl, an amine protecting group, phenyl or heteroaryl. 
     
     
         113 . (canceled) 
     
     
         114 . The conjugate of  claim 82 , wherein the conjugate is represented by one of the following formulae: 
       
         
           
           
               
               
           
         
       
     
     
         115 . The conjugate of  claim 82 , wherein L CB  is a linker bearing —C(═O)— group that is covalently linked to the cell-binding agent. 
     
     
         116 . The conjugate of  claim 82 , wherein (i) L CB  is represented by any one of the following formulae:
   —C(═O)—NR 5a —R x1 —S Z   s1 -  (L3a′),
     —NR 5a —C(═O)—R x2 —S Z   s1 -  (L3b′)
     —C(═O)—NR 5a —R x3 - J   CB -  (L3c′), or
     —NR 5a —P 2 —C(═O)—R x4 - J   CB -  (L3d′),
   
       wherein:
 R 5a  is H or (C 1 -C 3 )alkyl; 
 R x1 , R x2 , R x3  and R x4  are each independently a (C 1 -C 10 )alkyl, a (C 3 -C 8 )cycloalkyl, an aryl or a heteroaryl, 
 P 2  is an amino acid residue or a peptide containing between 2 to 20 amino acid residues; 
 J CB  is —C(═O)— covalently linked to the CBA; 
 R a  is —OH, —Cl, —O(C 1 -C 6 )alkyl; 
 Z s1  is a bifunctional linker that is covalently linked to the cell-binding agent; and 
 R e  is a (C 1 -C 6 )alkyl or is selected from phenyl, nitrophenyl (e.g., 2 or 4-nitrophenyl), dinitrophenyl (e.g., 2,4-dinitrophenyl), carboxynitrophenyl (e.g., 3-carboxy-4-nitrophenyl), pyridyl or nitropyridyl (e.g., 4-nitropyridyl); and 
 R e1  is a (C 1 -C 6 )alkyl; or 
 
       (ii) Len is represented by the following formula:
   —C(═O)—NR 5a —R x1 —S- Z   c1a   (L4a′)
 
   —NR 5a —C(═O)—R x2 —S Z   c1a   (L4b′)
 
   —C(═O)—NR 5a —R x3 —C(═O)- Z   c2a   (L4c′), or
 
   —NR 5a —P 2 —C(═O)—R x4 —C(═O)- Z   c2a   (L4d′),
 
 
       wherein:
 Z c1a  is 
 
       
         
           
           
               
               
           
         
         Z c2a  is 
       
       
         
           
           
               
               
           
         
         s9 is the site connected to Ar 3  or Ar 2  when n3 is 0, and s10 is the site connected to the CBA through the thiol group on a cysteine; 
         V is —C(═O—NR 9 -, or —NR 9 —C(═O)—; 
         Q is —H, charged substituent or an ionizable group; 
         R 9 , R 10 , R 11 , R 12 , R 13 , R 19 , R 20 , R 21  and R 22  for each occurrence, are independently —H or a (C 1 -C 4 )alkyl; 
         r1 and r2 are each independently 0 or an inter between 1 and 10; 
         q1 and q2 are each independently 0 or an integer between 1 and 10; 
         R h  is —H or a (C 1 -C 3 )alkyl; 
         R 19  and R 20 , for each occurrence, are independently —H or a (C 1 -C 4 )alkyl; 
         S1 is an integer between 1 and 10; 
         P 3  is an amino acid residue or a peptide residue containing 2 to 5 amino acid residues; or 
       
       (iii) Len is represented by the following formula:
   —C(═O)—NR 5a —R x1 —S- Z   s1 - J   CB   s1   (L5a′)
 
   —NR 5a —C(═O)—R x2 —S Z   s1 - J   CB   s1   (L5b′)
 
   —C(═O)—NR 5a —R x3 - Z   a2 —R x3′ - J   CB   s1   (L5c′)
 
   —NR 5a —R x3 —C(═O)—R x4 - Z   a2 —R x4′ - J   CB   s1   (L5c1′), or
 
   —NR 5a —P 3 —C(═O)—R x4 - Z   a2 —R x4′ - J   CB   s1   (L5d′),
 
 
       wherein:
 R x1 , R x2 , R x3 , R x4  and R x4′  are each independently a (C 1 -C 6 )alkyl; 
 Z s1  is 
 
       
         
           
           
               
               
           
         
         s9 is the site covalently linked to the group J CB ′; 
         s10 is the site covalently linked to the —S— group in formula (L5a) or (L5b); 
         Z a1  is absent, —C(═O)—NR 9 -, or —NR 9 —C(═O)—; 
         R 9  is —H or a (C 1 -C 3 )alkyl; 
         Q is H, a charged substituent or an ionizable group; 
         R a1 , R a2 , R a3 , R a4  for each occurrence, are independently H or C 1 -C 3 )alkyl; and 
         qs1 and rs1 are each independently an integer from 0 to 10, provided that qs1l and rs1 are not both 0 
         Z a2  is absent, —C(═O)—NR 9 -, or —NR 9 —C(═O)—; 
         R 9  is —H or a C 1 -C 3 )alkyl; 
         J CB   s1  is a moiety formed by reacting an aldehyde reactive group with an aldehyde group located on the CBA. 
       
     
     
         117 - 129 . (canceled) 
     
     
         130 . The conjugate of  claim 82 , wherein the conjugate is any one of the following: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       or a pharmaceutically acceptable salt thereof, wherein 
       
         
           
           
               
               
           
         
       
       represents the cell-binding agent covalently linked to the cytotoxic agent via the ε-amino group of a lysine, the thiol group of a lysine, or an aldehyde group located on the CBA;
 the double line   between N and C represents a single bond or a double bond, provided that when it is a double bond X is absent and Y is —H and when it is a single bond, X is —H ad Y is —SO 3 H; 
 Z 101  is represented by one of the following formulae: 
 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         q is an integer from 1 to 5; 
         n′ is an integer from 2 to 6; 
         w c  is 1 or 2; 
         w s  is 1 or 2; and 
         w is an integer from 1 to 10. 
       
     
     
         131 - 154 . (canceled) 
     
     
         155 . The conjugate of  claim 82 , wherein
 the double line   between N and C represents a double bond, X is absent and Y is —H; or   the double line  between N and C represents a single bond, X is —H and Y is —SO 3 H or —SO 3 Na.   
     
     
         156 . (canceled) 
     
     
         157 . A pharmaceutical composition comprising a conjugate of  claim 82 , and a pharmaceutically acceptable carrier. 
     
     
         158 . A method of inhibiting abnormal cell growth or treating a proliferative disorder, an autoimmune disorder, destructive bone disorder, infectious disease, viral disease, fibrotic disease, neurodegenerative disorder, pancreatitis or kidney disease in a mammal, comprising administering to said mammal a therapeutically effective amount of a conjugate of  claim 82 , and optionally, a chemotherapeutic agent. 
     
     
         159 . The method of  claim 158 , wherein the method is for treating a condition selected from cancer, rheumatoid arthritis, multiple sclerosis, graft versus host disease (GVHD), transplant rejection, lupus, myositis, infection, and immune deficiency, and optionally, the cancer is ovarian cancer, pancreatic cancer, cervical cancer, melanoma, lung cancer (e.g. non small-cell lung cancer and small-cell lung cancer), colorectal cancer, breast cancer (e.g. triple negative breast cancer (TNBC)), gastric cancer, squamous cell carcinoma of the head and neck, prostate cancer, endometrial cancer, sarcoma, multiple myeloma, head and neck cancer, blastic plasmacytoid (MDS), peritoneal cancer, or leukemia (e.g. acute myeloid leukemia (AML), acute monocytic ALL), chronic lymphocytic leukemia (CLL), and chronic myeloid leukemia (CML)). 
     
     
         160 - 161 . (canceled)